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Flow Cytometry-based Purification of S. cerevisiae Zygotes
JoVE 杂志
生物学
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JoVE 杂志 生物学
Flow Cytometry-based Purification of S. cerevisiae Zygotes

Flow Cytometry-based Purification of S. cerevisiae Zygotes

DOI:
10.3791/4197-v

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15:09 min

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September 21, 2012

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, , , ,
1Department of Pathology,Case Western Reserve University School of Medicine, 2Cell Biology Program,Case Western Reserve University School of Medicine, 3Case Comprehensive Cancer Center,Case Western Reserve University School of Medicine

Chapters

  • 00:05Title
  • 02:03Haploid Cell Growth
  • 03:08Conducting a Cross
  • 04:43Harvesting Preparations
  • 05:57Flow Cytometry
  • 09:01Preparation for DeltaVision Microscopy
  • 11:05Representative Analysis of Zygote Purification
  • 14:27Conclusion

Summary

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To purify zygotes of S. cerevisiae, haploid cells of opposite mating type were engineered to express red or green fluorescent proteins, co-incubated to allow zygote formation, and fractionated using a flow cytometry-based protocol. The highly-enriched fraction enables subsequent "-omic" studies, recovery of initial progeny, and systematic investigation of zygote morphogenesis.

Tags

Flow Cytometry-based PurificationS. Cerevisiae ZygotesHaploid CellsDiploid StatesMating TypeStable DiploidsMitotic DivisionsZygote FormationSoluble Mating FactorsReceptor-mediated Signaling CascadesCell-cell FusionModel For Progenitor Or Stem Cell FunctionBiochemistry Of Zygote FormationPurification Of Yeast ZygotesSedimentation PropertiesHypertonic Sorbitol

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